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CYP1A2 Assay Systems

The P450-Glo™ CYP450 Assays provide a homogeneous, luminescent method for measuring cytochrome P450 activity. The assays are designed to measure the activities of P450s from recombinant and native sources and for testing the effects of analytes such as drugs and new chemical entities on P450 activities. These luminescent assays exhibit exquisite sensitivity, low background signals and broad dynamic range.

P450-Glo™ Assays employ luminogenic P450 substrates that are derivatives of beetle luciferin, a substrate for luciferase enzymes. The derivatives are not substrates for luciferase but are converted by P4...

The P450-Glo™ CYP450 Assays provide a homogeneous, luminescent method for measuring cytochrome P450 activity. The assays are designed to measure the activities of P450s from recombinant and native sources and for testing the effects of analytes such as drugs and new chemical entities on P450 activities. These luminescent assays exhibit exquisite sensitivity, low background signals and broad dynamic range.

P450-Glo™ Assays employ luminogenic P450 substrates that are derivatives of beetle luciferin, a substrate for luciferase enzymes. The derivatives are not substrates for luciferase but are converted by P450s to luciferin, which in turn reacts with luciferase to produce light that is directly proportional to the activity of the P450.

The P450-Glo™ Assays generate a "glow-type" luminescent signal, produced using derivatized luciferins as P450 substrates and a recombinant stabilized luciferase (Ultra-Glo™ Luciferase) coupled with a proprietary buffer system. The half-life of the luminescent output is greater than two hours, eliminating the need for luminometers with injectors and allowing for batch plate processing. The formulation also minimizes the incidence of false positives due to inhibition of luciferase by analytes when screening for cytochrome P450 inhibitors.

The P450-Glo™ CYP1A2 Induction/Inhibition Assay contains a new substrate for cytochrome 1A2 that is very well suited for all applications involving human CYP1A2 and is the best substrate available for cell-based applications. The substrate, Luciferin-1A2, is readily taken up by cells and converted into a luciferin precursor by CYP1A2. The luciferin precursor is rapidly converted into luciferin following the addition of D-cysteine. After addition of luciferin-1A2 to cells, the assay can be completed in approximately one hour. The low background and high signal-to-noise ratio produced using Luciferin-1A2 means less starting material is required.

The P450-Glo™ CYP1A2 Screening System provides a complete set of reagents for performing luminescent CYP1A2 assays using recombinantly expressed CYP1A2. The system includes a membrane preparation containing recombinant human cytochrome 1A2, a luminogenic cytochrome P450 substrate (luciferin-ME), an NADPH Regeneration System, reaction buffer, Luciferin Detection Reagent and Luciferin-Free Water. The membranes are prepared from baculovirus-infected insect cells and contain human cytochrome P450 and P450 reductase. The P450-Glo™ Screening System also contains a membrane fraction devoid of cytochrome P450 activity as a negative control. The assay is ideal for testing the effects of drugs and new chemical entities on cytochrome P450 enzyme activities.

Excellent Selectivity:The P450-Glo™ CYP1A2 Induction/Inhibition Assay contains Luciferin-1A2, a pro-luciferin substrate that is more selective for CYP1A2, which makes it ideal for use in cell-based applications.

This product is available through the Promega Helix onsite stocking program in a –20°C Helix Freezer. The program offers numerous convenient solutions to meet your lab's needs. Helix Freezers are available in two sizes: 5.7 cubic ft. or 9.7 cubic ft.

This product is available through the Promega Helix onsite stocking program in a –20°C Helix Freezer. The program offers numerous convenient solutions to meet your lab's needs. Helix Freezers are available in two sizes: 5.7 cubic ft. or 9.7 cubic ft.

Storage Conditions

Store the CYP1A2 membranes at –70°C. Cytochrome P450 may lose activity with repeated freeze-thaw cycles. Avoid multiple freeze-thaw cycles by dispensing the CYP1A2 membranes into single-use aliquots (e.g., 50μl for 96 reactions). Store aliquots at –70°C. All other components can be stored at –20°C or –70°C and protected from light.

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